Functional Characterization of POFUT1 Variants Associated with Colorectal Cancer
| Autor: | Marlène Deschuyter, Sébastien Legardinier, Florian Pennarubia, Abderrahman Maftah, Emilie Pinault |
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| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
Cancer Research POFUT1 biology EGF-like domain Colorectal cancer Notch signaling pathway colorectal cancer medicine.disease lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens mutations lcsh:RC254-282 Article Targeted mass spectrometry Oncology chemistry Glycosyltransferase medicine biology.protein Cancer research Missense mutation Receptor Glycoprotein |
| Zdroj: | Cancers Cancers, Vol 12, Iss 1430, p 1430 (2020) Volume 12 Issue 6 |
| ISSN: | 2072-6694 |
| Popis: | Background: Protein O-fucosyltransferase 1 (POFUT1) overexpression, which is observed in many cancers such as colorectal cancer (CRC), leads to a NOTCH signaling dysregulation associated with the tumoral process. In rare CRC cases, with no POFUT1 overexpression, seven missense mutations were found in human POFUT1. Methods: Recombinant secreted forms of human WT POFUT1 and its seven mutated counterparts were produced and purified. Their O-fucosyltransferase activities were assayed in vitro using a chemo-enzymatic approach with azido-labeled GDP-fucose as a donor substrate and NOTCH1 EGF-LD26, produced in E. coli periplasm, as a relevant acceptor substrate. Targeted mass spectrometry (MS) was carried out to quantify the O-fucosyltransferase ability of all POFUT1 proteins. Findings: MS analyses showed a significantly higher O-fucosyltransferase activity of six POFUT1 variants (R43H, Y73C, T115A, I343V, D348N, and R364W) compared to WT POFUT1. Interpretation: This study provides insights on the possible involvement of these seven missense mutations in colorectal tumors. The hyperactive forms could lead to an increased O-fucosylation of POFUT1 protein targets such as NOTCH receptors in CRC patients, thereby leading to a NOTCH signaling dysregulation. It is the first demonstration of gain-of-function mutations for this crucial glycosyltransferase, modulating NOTCH activity, as well as that of other potential glycoproteins. |
| Databáze: | OpenAIRE |
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